ANTIBODY-RESPONSE TO BRUCELLA-MELITENSIS OUTER-MEMBRANE ANTIGENS IN NATURALLY INFECTED AND REV1 VACCINATED SHEEP

Sera from Brucella infected and B. melitensis Rev1 vaccinated sheep were analysed by immunoblotting using the cell envelope fraction (CEF) of B. melitensis B115. The CEF of B. melitensis B115 was analysed using a bank of monoclonal antibodies. The fraction consisted mainly of S-LPS like molecules, R-LPS and outer membrane proteins (OMPs) of molecular masses of 10, 16.5, 19, 25-27, 31-34, 36-38, 73 and 89 kDa. Immunoblot analysis indicates that the antibody response in infected sheep was mainly directed against the major OMPs of 25-27, 31-34, 36-38 kDa, against 55 to 62, 70-73 and 89 to 94 kDa proteins associated with the CEF and, against S-LPS like molecules. Some infected sheep reacted with antigens of molecular mass lower than 20 kDa. Sera from vaccinated sheep reacted only with OMPs of 36-38, 60, 70-73 and 89 kDa. The major 25-27 and 31-34 kDa OMPs and proteins below 20 kDa were only detected by the sera of infected sheep. These differences may be due to the persistence of the field infection also reflected by the fact that antibody response against O-polysaccharide (O-PS), as measured by enzyme-linked immunosorbent assay (ELISA), was more intense in infected sheep than in vaccinated ones. These results also indicate that these OMPs could be useful to differentiate B. melitensis infection from B. melitensis Rev.1 vaccination in sheep.